Development of selective HDAC6 inhibitors to improve cancer immunotherapy

开发选择性 HDAC6 抑制剂以改善癌症免疫治疗

基本信息

批准号：
10337269
负责人：
Cyril Barinka
金额：
$ 10.43万
依托单位：
GEORGE WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-02-01 至 2023-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10337269
关键词：
Address Adjuvant Advanced Malignant Neoplasm Anti-Inflammatory Agents Antineoplastic Agents Attention Autoimmunity Biological Assay Blocking Antibodies CD8B1 gene CTLA4 gene Cardiac Cell Therapy Cells Characteristics Chronic Clinical Combination immunotherapy Combined Modality Therapy Cytotoxic agent Deacetylase Development Distant Down-Regulation Effectiveness Epigenetic Process Event Generations Genetic Goals Graft Rejection Grant HDAC6 gene Histone Deacetylase Histone Deacetylase Inhibitor Immune Immune system Immunity Immunologic Surveillance Immunologics Immunotherapeutic agent In Vitro In complete remission Infiltration Inflammation Mediators Inflammatory Interleukin-10 Isoenzymes Knowledge Lead Life Expectancy Lung Malignant Neoplasms Measures Mediating Mediator of activation protein Metabolic Metastatic Melanoma Methods Modality Modeling Molecular Target Mus Neoplasm Metastasis Neurodegenerative Disorders Patients Pharmaceutical Chemistry Pharmaceutical Preparations Pharmacology Phenotype Population Positioning Attribute Process Property Proteins Radiation therapy Rattus Regulation Reporting Research Resistance Role Surface Testing Toxic effect Translating Treatment Efficacy Tubulin Tumor Antigens Tumor Immunity Tumor-associated macrophages Validation Vorinostat Work anti-PD-1 anti-tumor immune response base cancer cell cancer immunotherapy cancer therapy cytokine cytotoxic cytotoxicity design genotoxicity immune checkpoint immune checkpoint blockade immune-related adverse events immunoregulation improved in vivo inhibitor innovation interest macrophage melanoma model design molecular modeling neoplastic cell novel offspring patient derived xenograft model pre-clinical programmed cell death ligand 1 programmed cell death protein 1 rational design receptor restoration scale up side effect targeted agent translational study tumor tumor growth

项目摘要

Project Summary The median survival for metastatic melanoma is approximately 8–16 months, and few therapies offer a significant improvement in overall survival. However, immunotherapeutic strategies that abrogate immunologic checkpoints or improve immunosurveillance have shown promise, especially in melanoma. We have found that both genetic abrogation and pharmacologic inhibition of HDAC6 leads to a decreased infiltration of pro-tumoral tumor-associated macrophages and downregulation of immunosuppressive mediators. These effects translated into a pronounced delay of in vivo melanoma tumor growth, which is, at least in part, dependent on intact immunity, as evidenced by the restoration of tumor growth after CD4+ and CD8+ depletion. Our findings demonstrate a significant immunoregulatory role of HDAC6 in melanoma, providing a rationale for the use of selective HDAC6is to improve antitumor immunity. We are most interested in identifying HDAC6is that are best able to reduce the pro-tumoral phenotype of tumor-associated macrophages and decrease the expression of immunosuppressive surface molecules such as PD-L1 and PD-L2 while showing little cytotoxicity on their own. Our goal is thus to design, synthesize, and test new HDAC6i both in vitro and in vivo for use in the treatment of melanoma and other malignancies. The aims to be accomplished under this grant are as follows: 1. Using molecular modeling, design, and synthesize ~50 new HDAC6is per year, for the first three years and test these for HDAC isozyme selectivity; the last two years of the grant will focus on compound scale-up, preclinical translational studies, and advanced ADMET testing. 2. Evaluate compounds that have IC50 values of <50nM and at least 400-fold HDAC6 selectivity using in vitro melanoma models to measure acetylated tubulin, pSTAT3, PD-L1 levels, and cytotoxicity. For compounds found to decrease levels of pSTAT3 and PD-L1, while increasing acetylated tubulin, conduct early-stage ADMET studies (2 – 3/year) using a CRO, and if needed, modify the compounds using principles of medicinal chemistry and molecular modeling based methods to increase the compound’s drug-like character. 3. Lastly, test the best compounds for their ability to delay tumor growth in syngeneic murine melanoma models, both as stand-alone and in combination with PD-1 blocking antibodies. Additionally, we will evaluate our best candidates using humanized PDX models and as a cell therapy pre-treating macrophages to avoid their switch to M2-like protumoral phenotype. Those candidates showing the best in vivo efficacy will be sent for advanced ADMET, including chronic toxicity studies, cardiac activity, and pulmonary activity in rats. Our proposed work is significant as it helps to fill this critical knowledge gap and thus moves epigenetic- based combination immunotherapies to a new level. The concept to be validated is novel as it significantly deviates from the classical cytotoxic role of HDAC inhibitors as anti-cancer agents.

项目概要转移性黑色素瘤的中位生存期约为 8-16 个月，很少有疗法能提供长期有效的治疗。然而，消除免疫学的免疫治疗策略显着改善了总体生存率。检查点或改善免疫监视已显示出希望，特别是在黑色素瘤中。我们发现 HDAC6 的基因消除和药物抑制都会导致促肿瘤肿瘤相关巨噬细胞的浸润减少以及这些作用转化为体内黑色素瘤肿瘤的显着延迟。生长，至少部分依赖于完整的免疫力，肿瘤的恢复就证明了这一点 CD4+ 和 CD8+ 耗尽后的生长。我们的研究结果证明了其显着的免疫调节作用。 HDAC6 在黑色素瘤中的存在，为使用选择性 HDAC6 来提高抗肿瘤免疫力提供了理论依据。我们最感兴趣的是确定最能减少促肿瘤表型的 HDAC6 肿瘤相关巨噬细胞并减少免疫抑制表面分子的表达，例如与 PD-L1 和 PD-L2 一样，同时其本身几乎没有细胞毒性。因此，我们的目标是在体外和体内设计、合成和测试新的 HDAC6i，以用于黑色素瘤和其他恶性肿瘤的治疗这笔赠款要实现的目标如下： 1. 前三年每年使用分子建模、设计和合成约 50 个新的 HDAC6 测试这些 HDAC 同工酶的选择性；资助的最后两年将重点关注化合物的放大，临床前转化研究和高级 ADMET 测试。 2. 使用体外评估IC50值<50nM且HDAC6选择性至少400倍的化合物用于测量乙酰化微管蛋白、pSTAT3、PD-L1 水平和化合物细胞毒性的黑色素瘤模型。发现降低 pSTAT3 和 PD-L1 水平，同时增加乙酰化微管蛋白，进行早期阶段使用 CRO 进行 ADMET 研究（2 – 3/年），如果需要，使用药物原理修改化合物基于化学和分子建模的方法来增加化合物的药物样特征。 3. 最后，测试最佳化合物延缓同基因小鼠黑色素瘤肿瘤生长的能力模型，无论是独立的还是与 PD-1 阻断抗体组合。此外，我们将评估。我们最好的候选者使用人源化 PDX 模型并作为细胞疗法预处理巨噬细胞以避免那些表现出最佳体内功效的候选者将被发送到 M2 样原肿瘤表型。用于高级 ADMET，包括慢性毒性研究、大鼠心脏活动和肺活动。我们提出的工作意义重大，因为它有助于填补这一关键的知识空白，从而推动表观遗传- 基于联合免疫疗法的新水平的验证概念是新颖的，因为它显着。偏离了 HDAC 抑制剂作为抗癌药物的经典细胞毒作用。